The goal of this research is to develop a novel separation media technology to replace conventional polyacrylamide gels in electrophoresis. The new sieving matrices will employ pre-polymerized acrylamide-based polymers capable of thermo-reversible gelation similar to agarose. As a result, these gel matrices can be pre-formulated and supplied to users as a powder or solution that is simply heated and poured into standard horizontal and vertical electrophoresis units. By eliminating end-user involvement in the synthesis of polyacrylamide gels, significant enhancements in electrophoresis efficiency, safety and reproducibility will be achieved, while retaining the exceptional resolving power of traditional polyacrylamide. The Phase I objectives are to: (I) Develop the basic formulation requirements for optimizing gel physical properties (e.g., strength, clarity and rheology), (2) Develop specific gel formulations for use in dsDNA analysis, e.g., for PCR product analysis, mapping, etc., and (3) Develop gel formulations for mutation screening by heteroduplex and SSCP analysis. Phase II will leverage the Phase I research by extending the range of applications to include native and denaturing protein electrophoresis. PROPOSED COMMERCIAL APPLICATION: The electrophoresis matrices developed in this research will have immediate commercial utility as safe and convenient substitutes for polyacrylamide and related acrylamide-based gels. By eliminating the labor-intensive and often unreliable preparation of polyacrylamide gels starting from toxic monomers, the new matrices will provide important benefits to end-users.